14IBCAT1FEASIBILITY - Engineering a Nano-factory for Peptide Synthesis

Peptides are short chains of simple chemical building blocks called amino acids. They are involved in numerous key biological processes including acting as toxins, pigments, drugs and hormones. They also control many of the most important cellular functions in animals, plants and man, and are used as tools in scientific research. There is considerable worldwide interest in developing new methods of producing peptides in sufficient quantity and of sufficient quality for use as pharmaceuticals, agrochemicals or research tools. The current favoured method for manufacturing peptides involves using chemical agents to fuse together the amino acid building blocks that form them. This approach is time-consuming, expensive, generates toxic waste products, and cannot be applied to many unusual but valuable peptides. In this project we will develop a new method of producing peptides which is not limited by these major problems. We will use an engineered peptide 'nano-factory', which when introduced into bacteria allows them to produce significant quantities of high value 'difficult' peptides without any of the problems or complexities associated with chemical synthesis.

The global custom research peptide manufacturing market is currently valued at approximately £800M per annum. The future challenges facing this industry revolve around developing tractable routes to producing increasingly complex peptides, increasing peptide synthesis capability, and a requirement for new tools that facilitate access to historically intractable high value targets, e.g. amyloids, anti-microbial peptides, hydrophobic peptides and isotopically-labelled peptides. We have developed a bio-inspired nano-scale device, based on a ubiquitous bacterial chaperone, that sequesters and stabilises peptides during their biosynthesis, protecting them from proteolytic action and maintaining them in a fully folded and, where appropriate, a functional state. The objectives of this proposal are (i) to rationally re-engineer our prototype device to produce functionally-enhanced variants with broader utility, (ii) to integrate this technology into optimised microbial and cell-free expression platforms, and (iii) develop methods for efficient extraction and purification of the peptides following recombinant production, with a view to optimising peptide purity. Further preliminary investigations will take place to scale-up production with a view to the development of rapid methodologies for commercial production of pure peptides. The development of a ubiquitous method for recombinant protein production, even of peptides that are difficult to synthesise by traditional solid-state chemical methods, will provide great benefits.

As described in proposal submitted to IUK.